Specific detection of Salmonella serogroup D1 by polymerase chain reaction(PCR) for sefA gene

SefA 유전자 PCR에 의한 Salmonella serogroup D1의 특이적 검출

  • Jun, Moo-hyung (College of Veterinary Medicine, Chungnam National University) ;
  • Kim, Tae-joong (College of Veterinary Medicine, Chungnam National University) ;
  • Chang, Kyung-soo (College of Veterinary Medicine, Chungnam National University) ;
  • Kang, Kyong-im (College of Veterinary Medicine, Chungnam National University) ;
  • Kim, Kui-hyun (College of Veterinary Medicine, Chungnam National University) ;
  • Kim, Ki-seok (National Veterinary Research and Quarantine Services) ;
  • Yoo, Sang-sik (Taejon City Institute of Health and Environment) ;
  • Kim, Hyun-soo (College of Veterinary Medicine, Chungnam National University) ;
  • Shin, Kwang-soon (College of Veterinary Medicine, Chungnam National University) ;
  • Kim, Chul-joong (College of Veterinary Medicine, Chungnam National University)
  • Received : 1999.03.03
  • Published : 1999.06.22

Abstract

Sal enteritidis thin fimbriae, SEF14, were found to be restricted to the predominantly poultry-associated members of the Salmonella serogroup D1 that are considered as the important pathogens in poultry industry. SefA together with sefB and sefC encode the proteins involved in SEF14 biosynthesis. In order to develop the rapid and specific detection methods for Salmonella serogroup D1, a PCR technique for the amplification of sefA gene was established, and its specificity and sensitivity were investigated with various microorganisms. The bacterial genomic DNA was extracted by colony-picking and rapid boiled-lysate technique. In comparison of Sef I and Sef II primers used in the PCR, Sef I primer for sefA gene of 513bp showed higher specificity than that of Sef II. The established PCR was as sensitive as to detect 1pg of Sal enteritidis DNA. When 73 strains in 28 genera including the reference strains and the field isolates of various Salmonella serotypes, Bacillus subtilis, Bordetella bronchisepdca, E coli, Listeria spp., Micrococcus luteus, Rhodococcus equi, Staphylococcus spp., Streptococcus spp., Vibrio parahemolyticus, Yersinia spp. were studied, the established PCR yielded specifically positive results with only Salmonella serogroup D1. The results suggested that the PCR for sefA gene could be a potential candidate among the specific detection methods for Salmonella serogroup D1.

Acknowledgement

Supported by : 농림기술관리센터